FR3004810A1 - USE OF BARRIER BIOMARKERS FOR EVALUATION OF THE EFFECTIVENESS OF ASSETS. - Google Patents

Abstract

The subject of the invention is a method for evaluating the efficacy of an active agent selected from avocado sugars in C7, also called avocado perseoses, for preventing or treating a deficiency of the cutaneous barrier, said method comprising the determination of level of expression and / or activation of at least one biological marker, wherein said biological marker is selected from the markers of epidermal maturation, lipid barrier markers, markers of water regulation, markers of the regulation of stratum granulosum.

Description

The present invention relates to the field of theranostics, also sometimes called theragnostics, that is to say to the evaluation of the effectiveness of active agents. or compounds in the treatment of subjects, particularly with a view to adapting their treatment individually for personalized medicine. More specifically, the invention belongs to the field of dermatology, and relates to methods for evaluating the effectiveness of an active agent selected from C7 sugars and derivatives of formula (I), in the prevention and / or treatment at least one deficiency of the cutaneous barrier of a subject. The skin protects the body against external factors of physical constraints (mechanical, thermal factors and UV rays, in particular), chemical (surfactants, prolonged exposure to water, solvents, etc.) and environmental factors. In addition, the skin, as a barrier, protects the body against the loss of essential ions, water and serum proteins. The epidermis is the most superficial layer of the skin and ensures the impermeability and resistance of the skin. Four distinct cell layers, a basal layer (stratum basalis), a spinous layer (stratum spinosum), a granular layer (stratum granulosum), and a horny layer (stratum comeum) can be identified in the epidermis. Among them, it is in particular the stratum corneum (stratum comeum) and the granular layer (stratum granulosum), that is to say the outermost layers of the epidermis, which ensure the barrier function of the skin. The stratum corneum is usually composed of cornified keratinocytes, also called corneocytes. Cornified keratinocytes are cells that have lost their cell nucleus that result from a continuous process of keratinocyte differentiation, also known as keratinocyte maturation or epidermal maturation. The cornified keratinocytes are connected to each other by means of symbols, and are organized into an array of cells contiguous to each other and forming flexible and resistant lamellae. Between the cornified keratinocytes, the intercellular domain is rich in lipids, in particular those of the ceranid family. In addition, localized in the stratum corneum, are "natural moisturizing factors" (NMN), which maintain the degree of hydration of the skin at an optimal level.

This barrier made by the stratum corneum is not absolute. Indeed, there is a transepidernnic loss of water, or insensible loss of water, which is minimal, but which can increase in certain situations of deficiency of the barrier function. The barrier function of the skin evolves with the age of the subject, and may be incomplete or even deficient at certain ages of life. It is known for example that the skin of the child is not mature, and that its barrier function evolves with its maturation. In particular, the barrier of children's skin is only incompletely functional in comparison with adult skin.

Fluhr et al (Br J Dermatol 166 (3): 483-90, 2012) have shown that the water content of the stratum corneum is lower at birth (children aged 1 to 15 days) than in adults . Premature babies in particular show an increase in insensible loss of water (insensible water losses are multiplied by about 10 in premature infants) and a high sensitivity to infections, indicating the immaturity of the cutaneous barrier in this population of subjects. On the other hand, the skin of the elderly also shows a decrease in the cutaneous barrier, linked to the phenomenon of senescence, also called intrinsic aging (J. Lübbe, Swiss Medical Review, 62 (2472): 488-490, 2004) .

The skin barrier can also be weakened by alterations in its structure, which can result from both endogenous and exogenous factors. For example, it is estimated that infant acne, adolescent acne, rosacea or erythrocouperosis, psoriasis, seat dermatitis, atopic dermatitis, eczema, contact dermatitis, irritative dermatitis and in particular irritative dermatitis of the seat or diaper rash, allergic dermatitis, seborrheic dermatitis, dry skin, hyperreactivity of the skin, to mention only these pathologies, are linked to a deficiency of the cutaneous barrier. Active agents to prevent or restore the barrier function of the skin are already known in the art. In particular, the protective effect for the barrier function of C7 sugars and their derivatives, avocado perseose, has already been described (see for example WO 2005/105123). However, no biological marker has yet been known to measure the efficacy of said avocado perseosis during treatment in a particular subject. However, the effectiveness of a dermatological treatment may depend on the condition of the skin of the treated subject. As a result, it was not possible until now to adapt or adjust said treatment as a function of subject treated. The present inventors have, for the first time, identified markers that are specifically expressed or activated when the skin is treated with avocado perseose. In particular, they observed that some biological markers are more expressed or more active in avocado perseose-treated skin than in untreated skin. These markers can thus be used to evaluate the efficacy of avocado perseosis in preventing or treating a barrier deficiency in a subject. DETAILED DESCRIPTION According to a first aspect, the subject of the invention is a method for evaluating the efficacy of an active agent selected from avocado sugars in C7, also called avocado perseose, which will be defined later on for preventing or treating a cutaneous barrier deficiency, said method comprising determining the level of expression and / or activation of at least one biological marker.

It is understood that the assets of the invention are used in mammals, and preferably humans. More preferably, said compositions are used in children. "Subject" means, within the meaning of the present application, any human subject, irrespective of his age. Thus, the subject may be for example an adult or a child. By "child" is meant according to the invention an individual whose age is less than 16 years. Children included in the category of children according to the invention are therefore newborns, that is to say children whose age is between 0 and 1 month, infants, that is to say infants. children between 1 month and 2 years old, and children 2 years old or over, ie children between the ages of 2 and 16 years old. A "newborn", as we understand it here, may as well be born at term as it is premature.

For the avoidance of doubt, the term child used in this application without further specification must be understood in its most general sense, that is, referring to a person under 16 years of age. Thus, in the context of the present invention, children aged 2 to 16 are specifically referred to by the terms - children aged 2 or over.

An "adult" within the meaning of the present invention is a person who is not a child, that is, a person over 16 years of age. According to a particular embodiment, the subject is a child, preferably a newborn or an infant. According to a preferred embodiment, the subject of the invention is a method for evaluating the effectiveness of an active agent chosen from C7 sugars and derivatives of formula (I), CH2ORi Ra-C-Rb R30 H R40 HH OR6 H OR6 CH2OR7 (I) wherein Ra represents a hydrogen atom and Rb represents a -OR2 or CRaRb represents the CO radical; R1, R2, R3, R4, R5, R6 and R7 represent, independently of one another: - a hydrogen atom or - a radical - (CO) -R in which R represents a saturated or unsaturated hydrocarbon chain containing from 11 to 24 carbon atoms, optionally substituted with one or more substituents selected from the group consisting of hydroxy (-OH), ethoxy (-OC2H5) and -503M with M representing a hydrogen atom, an NH4 + ammonium ion or a metal ion; or a - (CO) -R 'radical in which R' represents a saturated or unsaturated hydrocarbon chain containing from 2 to 10 carbon atoms, optionally substituted with one or more substituents chosen from the group consisting of hydroxyl (-OH) radicals; ethoxy radicals (-OC2H5) and -503M group with M representing a hydrogen atom, an NH4 + ammonium ion or a metal ion; in the prevention and / or treatment of at least one skin barrier deficiency of a subject, said method comprising the steps of: a. measuring the level of expression and / or activation of at least one biological marker in a sample of cutaneous cells of the subject, characterized in that said biological marker is chosen from: the markers of the epidermic maturation, said marker of the epidermal ripening being preferably chosen from desnyline 1 and involucrin, or; the markers of the lipid barrier, said lipid barrier marker being preferably chosen from ceranides, in particular from ceranides 1 to 9, even more particularly ceranide 1, or; the markers of the water regulation, said marker of the water regulation being preferably chosen from filaggrin, PAD1, hyaluronic acid and transglutanninase 1, or; the markers of the regulation of the stratum granulosum, said regulation marker of the stratum granulosum being preferably chosen from claudines, in particular claudin 1; b. measuring the level of expression and / or activation of said biological marker in a sample of cutaneous reference cells; vs. comparing the levels of expression and / or activation obtained in step a) with expression and / or activation levels obtained in step b); d. evaluating the effectiveness of said asset based on the comparison of step b). As is well known to those skilled in the art, the barrier function of the skin corresponds to the ability of the skin to limit the exchanges between the external environment and the interior of the body, more particularly the diffusion of water. The integrity of the skin barrier function or the integrity of the skin barrier (as understood here, the two terms are synonymous) means that the cutaneous barrier is fully functional, that is to say that trade, and in particular the diffusion of water, is limited.

The integrity of the skin barrier can be assessed by measuring a large number of parameters. In particular, it is usual to determine the integrity of the skin barrier by measuring the insensible loss of water, also known as transepidermal water loss. Methods for measuring water-insensitive loss are well known to those skilled in the art and do not need to be described in detail here (see, for example, Taganni and K Kuchuchi, Diseases that affect barrier function. In Elias PM and Feingold KR editors, Skin Barrier, New York: Taylor and Francis, 2005. p447-468). Preferably, to measure the water-insensitive loss, an evaporinometer is used, which is composed of a probe which is placed 3 or 6 mm above the skin.

D-nannoheptulose, the first ketoheptose identified in 1916 by La Forge, of general formula (II) is a C7 sugar found in certain plants, in particular in alfalfa (Medicago sativa L.), avocado, fig (Ficus officinalis L.) in the stonecrop (Sedum spectabile Bor.) and primrose (Prirnula officinalis Jacq.). However, it is in the avocado, that one finds the most important contents in D-enannoheptulose. D-Mannoheptulose has already been used in therapeutic applications. For example, patent application WO 95/03809 discloses the use of D-amannoheptulose, as a glucokinase inhibitor, to inhibit the development of tumor cells and US 2003/0092669 discloses an oral dietary supplement comprising D - Annoheptulose, which reduces the insulin level and thus allows weight loss. Perseitol, a polyol form of D-aminotheptulose, of general formula (III) is also found in avocado, in particular in the fruit or in the nucleus of Persineitol, associated with a potassium ion, has been shown to inhibit the incorporation of leucine-3H into Ehrlich sarcomatous ascites tumor cells (Shibuya et al., Pure Appl. Chem., 71 (6): 1109-1113, 1999). The use of these sugars (perseitol and D-annenoheptulose) to treat alopecia has already been described (WO 2011/073281). The use of perseose avocado in the treatment of candidiasis and pityrosporosis has also been described (WO 2008/025847). It was finally shown that these sugars could be used to stimulate the synthesis of human beta-defensins (in particular HBD-2) (WO 2005/115421, WO 2005/105123). In particular, these sugars were already known to be effective in the prevention or treatment of pathologies related to a deficiency of the cutaneous barrier (see for example WO 2005/105123). On the other hand, until now, no biological markers whose expression or activation correlated with the use of these sugars in the treatment of skin barrier deficiencies were identified. However, the present inventors have identified such biological markers, which makes it possible to evaluate the effectiveness of said sugars in the prevention or treatment of deficiencies of the cutaneous barrier of a subject.

By the effectiveness of an active ingredient in the prevention of at least one deficiency of the cutaneous barrier of a subject, the term "application" means, within the meaning of the present application, the ability of said active ingredient to maintain the functional integrity of the cutaneous barrier in the body. subject. In other words, the effectiveness of an asset in preventing at least one deficiency of a subject's skin barrier corresponds to the ability of said asset to prevent the deterioration of the skin barrier function of said subject. By "efficacy of an active ingredient in the treatment of at least one deficiency of the cutaneous barrier of a subject", is meant within the meaning of the present application the ability of said active ingredient to restore the functional integrity of the cutaneous barrier in the body. subject. In other words, the effectiveness of an active ingredient in the treatment of at least one deficiency of a subject's skin barrier corresponds to the ability of said active agent to reverse the deterioration of the skin barrier function of said subject. By "biological marker" is meant in the sense of the present application a feature that is objectively measured and evaluated as an indicator of normal biological processes, pathogenic processes, or pharmacological responses to a therapeutic intervention. A biological marker therefore refers to a variety of different substances and parameters. For example, a biological marker may be a substance whose detection indicates a particular disease state (for example the presence of activated protein C as a marker of infection), or on the contrary a substance whose detection indicates a physiological state specific. The biological marker according to the invention is preferably a gene, the products of a gene such as its transcripts and the peptides derived from its transcripts, a lipid, a sugar or a metabolite.

According to one embodiment of the present invention, the biological marker is a gene, the products of a gene such as transcripts or peptides, a lipid, a sugar or a metabolite whose expression changes, in particular level expression, correlate with a physiological state of the cutaneous barrier.

According to one embodiment of the present invention, the biological marker is a gene, the products of a gene such as transcripts or peptides, a lipid, a sugar or a metabolite whose activation changes, in particular level activation, correlate with a physiological state of the cutaneous barrier. According to a particular embodiment, the biological marker is a peptide having an enzymatic activity. According to the invention, the biological marker is chosen from the markers of epidermal maturation, the markers of the lipid barrier, the markers of the water regulation, and the markers of the regulation of the stratum granulosum. The inventors have in particular demonstrated that a perseose treatment of avocado leads to an increase in the expression of key markers of epidermal maturation such as desnnylin and involucrine in skin models in vitro. The involucrine protein is expressed in the spinous-granular layers. It is the first precursor of the horny envelope which represents 5 to 15% of the horny envelope, and serves as a link also with the horny lipid envelope. The desnnylin protein is synthesized by keratinocytes and secreted at the stratum granulosum-stratum comeum interface. It is an indispensable component of corneodes, the junctions that bind corneocytes. The measurement of the expression of these markers in the subject treated with the active ingredient of the invention therefore makes it possible to measure that it is very effective in said subject, in particular on epidermal maturation. Preferably, the marker of epidermal maturation is chosen from desnnylin 1 and involucrine. For the purpose of the present invention, the biological marker desnnylin 1 comprises the human DSG1 gene (reference NCBI: GENE ID 1828), as well as the products of this gene. The products of the human DSG1 gene include the human DSG1 gene transcript and the human desnylinin 1 precursor peptide. For the purposes of the present invention, the transcript of the human DSG1 gene is the polypeptide whose sequence is for reference NCBI NM_001942.2. For the purposes of the invention, the term peptide precursor of the human desnoglein 1 protein is intended to mean the peptide whose peptide sequence is the reference sequence NCBI: NP_001933.2. For the purposes of the present invention, the involucrine biological marker comprises the human IVL gene (reference NCBI: Gene ID: 3713), as well as the products of this gene. The products of the human IVL gene include the human IVL gene transcript and human involucrine protein. For the purposes of the present invention, the transcript of the human IVL gene is the polypeptide whose sequence is for reference NCBI NM_005547.2. For the purposes of the invention, the term "human involucrin protein" means the protein whose peptide sequence is the reference sequence NCBI: NP_005538.2. The inventors have furthermore shown that avocetainin persearose in vitro, an increase in the expression of ceranides 1 to 9, in particular of ceranide 1. Cerannids are the major lipids of the lamellar bilayer and are therefore very important for the structure and functionality of the stratum comeum as a barrier. In particular, ceramide 1 is under-represented in dry, aged skin and also in many dermatoses such as atopic dermatitis, psoriasis, ichthyosis and acne. The measurement of these markers in the subject treated by perseose avocado thus offers a simple and solid tool to measure the effectiveness of the asset, and more particularly the effectiveness of the asset on the maintenance or restoration of the lipid barrier.

Preferably, the marker of the lipid barrier is chosen from ceranides. More preferably, the marker of the lipid barrier is chosen from ceranides 1 to 9. Even more preferentially, the marker of the lipid barrier is the ceranide 1. The filaggrin protein aggregates with the keratin fibers of the cytoskeleton, thus reducing the corneocytes in flattened disks, this intracellular network confers resistance and protection to stratunn corneunn. On the other hand, its degradation leads to the formation of the constituents of the NMF (Natural Moisturizing Factor), which is essential for the retention of water in corneocytes. . The degradation of filaggrin is provided by peptidylarginine deinninases (PADI) including PAD1 (Peptidyl Arginine Deinninase Type 1). Hyaluronic acid is found in the epidermis where it plays a role in the barrier function and hydration of the stratum corneum. The enzyme transglutaminase 1 participates in the establishment of the horny envelope by catalyzing the crosslinking of the protein involucrine. The measurement of at least one of these markers can therefore be used to estimate the effectiveness of the active agent on the hydric regulation of the stratum corneum. Preferably, the water regulation marker is chosen from filaggrin, PAD1 (Peptidyl Arginine Deinninase Type 1), hyaluronic acid and transglutaminase 1. For the purposes of the present invention, the biological marker filaggrin comprises the human FLG gene (reference NCBI: Gene ID: 2312), as well as the products of this gene. The products of the human FLG gene include human FLG gene transcript and human filaggrin protein. For the purposes of the present invention, the transcript of the human FLG gene is the polypeptide whose sequence is for reference NCBI NM_002016.1. For the purposes of the invention, the term "human filaggrin protein" means the protein whose peptide sequence is the reference sequence NCBI: NP_002007.1. For the purposes of the present invention, the biological marker PAD1 comprises the human PADI1 gene (reference NCBI: Gene ID: 29943), as well as the products of this gene. The products of the PADI1hunnain gene include the transcript of the human PADI1 gene and human protein-Peptidyl Arginine Deinninase Type 1, hereinafter referred to as human PAD1 protein. For the purposes of the present invention, the transcript of the human PADI1 gene is the polypeptide whose sequence is for reference NCBI NM_013358.2. For the purposes of the invention, the term "human PAD1 protein" refers to the protein whose peptide sequence is the reference sequence NCBI: NP_037490.2. For the purposes of the present invention, the transglutaminase 1 biological marker comprises the human TGM1 gene (reference NCBI: Gene ID: 7051), as well as the products of this gene. The products of the human TGM1 gene include the transcript of the human TGM1 gene and the human transglutaminase 1 protein. For the purposes of the present invention, the transcript of the human TGM1 gene is the polypeptide whose sequence is for reference NCBI NM_000359.2. For the purposes of the invention, the term "human transglutaminase 1 protein" refers to the protein whose peptide sequence is the reference sequence NCBI: NP_000350.1. In addition, it is known that the stratum granulosum plays a role in the proper functioning of the stratum comeum, and thus participates in the barrier function. The inventors have shown that, surprisingly, perseose of avocado makes it possible in vitro to increase the expression of the claudin1 protein. The claudin1 protein is a protein with 4 transnaninbranary domains belonging to a family of 24 members. It is a constituent of the tight junctions of stratum granulosum, which form a selective barrier controlling the paracellular transport of molecules, inflammatory cells. These tight junctions are also essential to limit water loss. Thus, by measuring the level of expression of markers of stratum granulosum regulation, it is possible to evaluate or verify the efficacy of avocado perseosis on a particular subject. Preferably, the regulatory marker of stratum granulosum is chosen from claudines. More preferably, the regulatory marker of the stratum granulosum is claudin 1. For the purposes of the present invention, the claudin 1 biological marker comprises human CLDN1 (reference NCBI: Gene ID: 9076), as well as the products of this gene. The products of the human CLDN1 gene include the human CLDN1 gene transcript and the human claudin 1 protein. For the purposes of the present invention, the transcript of the human CLDN1 gene is the polypeptide whose sequence is for reference NCBI NM_021101.4. For the purposes of the invention, the term "human claudin1 protein" means the protein whose peptide sequence is the reference sequence NCBI: NP_066924.1.

Those skilled in the art seeking to determine which class a gene or protein marker belongs to can easily consult the relevant scientific literature or refer to public databases such as, for example, those grouped on the website of the National Center for Biotechnology. Information (http://www.ncbi.nlnn.nih.gov/guide/). According to a particular embodiment, step a) comprises measuring the level of expression and / or activation of a combination of biological markers from a sample of cutaneous cells of the subject, characterized in that said marker combination comprises: at least one marker of epidermal maturation, said marker of epidermal maturation being preferably selected from desnnylin 1 and involucrin; at least one marker of the lipid barrier, said marker of the lipid barrier being preferably chosen from ceranides, in particular from ceranides 1 to 9, even more particularly ceranide 1; and at least one marker of the water regulation, said water regulation marker being preferably selected from filaggrin, PAD1, hyaluronic acid and transglutanninase 1; at least one stratum granulosum regulation marker, said stratum granulosum regulation marker being preferably selected from claudines, in particular claudin 1 Preferably, the combination of biological markers consists of desnnylin 1, involucrin, ceranide 1, filaggrin, PAD1, hyaluronic acid, transglutaminase 1 and claudin 1. When the biological marker is a gene, the terms "level of expression of the biological marker" refer to the level of synthesis of the products of this gene. More specifically, when the biological marker is a gene, the level of expression of said biological marker corresponds to the amount or concentration of transcripts of said gene and / or different isoforms of the proteins derived from said transcripts in the sample. When the biological marker is a lipid, a sugar, or a metabolite, the term "level of expression of the biological marker" refers to the level of synthesis of said lipid, said sugar, or said metabolite. More specifically, when the biological marker is a lipid, a sugar or a metabolite, the level of expression of said biological marker corresponds to the amount or the concentration of said lipid, said sugar or said metabolite in the sample. The terms "activation level of the biological marker" apply to biological markers for which biological activity, particularly enzymatic activity, can be measured. The person skilled in the art knows, for example, that certain peptides exhibit enzymatic activity. Those skilled in the art seeking to determine whether a biological marker of interest exhibits an activity, in particular enzymatic activity, can easily consult the relevant scientific literature or refer to public databases such as, for example, those grouped on the site. internet ExPASy, which lists enzymes in particular (http://enzyrne.expasy.org). According to the invention, the markers PAD1 and transglutaminase 1 are biological markers for which an enzymatic activity can be measured. When the biological marker is a biological marker for which enzymatic activity can be measured, the terms - level of activation of the biological marker "refer to the level of enzymatic activity of said marker. For the purposes of the present invention, the enzymatic activity of said label corresponds to the amount of substrate of said label catalyzed per unit of time thanks to a defined quantum of enzyme. The unit of enzymatic activity U is conventionally expressed as anol / nnin or kat (mmol / s). The unit of enzymatic activity U can also be expressed in mg / min when the substrate of the enzyme exists in the body in the form of a polymer, as is the case for example when the substrate is chosen from sugars or glycosaminoglycans.

By measuring the level of expression and / or activation of a combination of biological markers, it is understood within the meaning of the present application to measure the level of expression and / or activation of each of the markers of the combination. The expression of a gene can be measured for example at the nucleotide level, by measuring the amount of transcripts of said gene and their variants, and can also be measured for example at the protein level, for example by measuring the amount of one or more isofornnes proteins derived from said transcripts, and variants thereof. Thus, by measuring the level of expression of said gene, the term "measuring" means the quantity of product of the gene in its peptide form and / or in its nucleotide form. By "desnnhelin 1 in its peptide form" is meant according to the invention a peptide whose peptide sequence comprises the peptide sequence of the human desnoglein 1 protein (whose coding sequence is referenced CCDS11896.1), and / or its precursor peptide. . By "precursor peptide of human desnoglein 1 protein" is meant human prepro-desnnylin 1 (reference NCBI: NP_001933.2).

By nucleosine 1 in its nucleotide form "is meant according to the invention at least one polynucleotide whose sequence comprises the sequence of the human DSG1 gene transcript (reference NCBI: GENE ID 1828). For the purposes of the present invention, the transcript of the human DSG1 gene is the polypeptide whose sequence is for reference NCBI NM_001942.2.

By "involucrine in its peptide form" is meant according to the invention a peptide whose peptide sequence comprises the peptide sequence of the human involucrine protein (reference NCBI: NP_005538.2).

By -involucin in its nucleotide form is meant according to the invention at least one polynucleotide whose sequence comprises the sequence of the human IVL gene transcript (reference NCBI: Gene ID: 3713). For the purposes of the present invention, the transcript of the human IVL gene is the polypeptide whose sequence is for reference NCBI NM_005547.2.

Filaggrin in its peptide form is understood to mean, according to the invention, a peptide whose peptide sequence comprises the peptide sequence of the human filaggrin protein (reference NCBI: NP_002007.1).

In the nucleotide form, "filaggrin" is understood to mean at least one polynucleotide whose sequence comprises the transcript sequence of the human FLG gene (reference NCBI: Gene ID: 2312). For the purposes of the present invention, the transcript of the human FLG gene is the polypeptide whose sequence is for reference NCBI NM_002016.1. By "PAD1 in its peptide form" is meant according to the invention a peptide whose peptide sequence comprises the peptide sequence of the human protein (reference NP_037490.2).

By "PAD1 in its nucleotide form" is meant according to the invention at least one polynucleotide whose sequence comprises the sequence of the human PADI1 gene transcript (reference NCBI: Gene ID: 29943). For the purposes of the present invention, the transcript of the human PADI1 gene is the polypeptide whose sequence is for reference NCBI NM_013358.2. By "transglutaminase 1 in its peptide form" is meant according to the invention a peptide whose peptide sequence comprises the peptide sequence of the human transglutaminase 1 protein (reference NCBI: NP_000350.1).

By "transglutaminase 1 in its nucleotide form" is meant according to the invention at least one polynucleotide whose sequence comprises the sequence of the human TGM1 gene transcript (reference NCBI: Gene ID: 7051). For the purposes of the present invention, the transcript of the human TGM1 gene is the polypeptide whose sequence is for reference NCBI NM_000359.2. Peptide 1 in its peptide form is understood to mean, according to the invention, a peptide whose peptide sequence comprises the peptide sequence of the human claudin 1 protein (reference NCBI: NP_066924.1).

By claudin 1 in its nucleotide form is meant according to the invention at least one polynucleotide whose sequence comprises the sequence of the human CLDN1 gene transcript (reference NCBI: Gene ID: 9076). For the purposes of the present invention, the transcript of the human CLDN1 gene is the polypeptide whose sequence is for reference NCBI NM_021101.4. According to a preferred embodiment of the invention, the level of expression of the marker chosen from desnnylin 1, involucrine, or filaggrin corresponds to the level of expression of said marker in its peptide form. According to a preferred embodiment of the invention, the expression level of PAD1 corresponds to the level of expression of said marker in its nucleotide form.

In general, the measurement of the level of expression and / or activation of the biological marker according to the invention will be detected in vitro from a sample of cutaneous cells of the subject. For the purposes of this application, the term "cutaneous skin sample of the subject" means any sample containing cells of the skin obtained from the subject. The skin cell samples according to the invention therefore comprise both the fresh skin explants obtained directly from the subject, the cutaneous skin cell cultures in suspension, the nonlinear skin cell cultures, the bilayer skin cell cultures and the models. tissues, including reconstructed skin cultures and reconstructed mucosal cultures.

Since it is often difficult to work on fresh explants, it is particularly advantageous in the context of the present invention to use skin cell cultures. Advantageously, the cutaneous cells according to the invention comprise normal, healthy or pathological cells, or cells derived from lines. For example, skin cells cultured may be cells obtained from skin tissue explant. By "explant" or "skin expierant" is meant here a sample of cells or cutaneous tissue, which may be made for surgical purposes or for performing analyzes. In particular, an explant can be obtained during a surgical excision. By "resection" is meant here a surgical procedure consisting of cutting (excising) a more or less wide or deep part of the skin to treat an abnormality or an outgrowth. Excision is performed either to remove a cancerous tumor or suspected of being, or to treat a benign skin abnormality that is troublesome, whether for functional or aesthetic reasons. An excision within the meaning of the invention includes for example the skin samples obtained after plastic surgery (mammary plasty, abdominal, facelift, preputial sampling, otoplasty, that is to say, ear gluing, syndactyly or supernumerary finger, etc. .). An explant can also be obtained by biopsy. "Biopsy" means here a sample of cells or skin tissue made for analysis. Several types of biopsy procedures are known and practiced in the field. The most common types include (1) incisional biopsy, in which only a sample of the tissue is removed; (2) excisional biopsy (or surgical biopsy), which consists of the total removal of a tumor mass, thus making a therapeutic and diagnostic gesture, and (3) the needle biopsy, in which a tissue sample is taken with a needle, it can be wide or thin. Other types of biopsies exist, such as smear or curettage, and are also encompassed by the present invention. The cutaneous cells according to the invention comprise at least one type of cell usually present in the hypoderm, the dermis and / or the epidermis. These cells thus comprise, inter alia, keratinocytes, melanocytes, fibroblasts, adipocytes, endothelial cells, mast cells, Langerhans cells and / or Merkel cells. Preferably, the cutaneous cells according to the invention comprise at least keratinocytes and / or fibroblasts. More preferably, the skin cells according to the invention comprise keratinocytes and / or fibroblasts. It may be interesting for a person skilled in the art to evaluate the effectiveness of the active agents on cells of the subject subjected to certain stresses, such as, for example, the parts of the subject's skin subjected to solar exposures, or, more generally, UV irradiation. According to one embodiment of the invention, the cutaneous cell sample comprises cells which have been irradiated, preferably by UV. According to the invention, the cutaneous cell sample can be processed prior to the measurement of the expression of the biological marker, for example in order to extract from said cutaneous cell sample an RNnn sample or a protein sample. The sample of RNnn or protein can then be directly used to measure the expression of the marker. Preparation or extraction of nRNA or proteins from a cell or tissue sample are routine procedures well known to those skilled in the art. For each of the types of biological markers of the invention, numerous methods are available to those skilled in the art for measuring the level of expression and / or activation of said biological marker. When the biomarker is a gene, and the level of expression of the marker is measured at the nucleotide level, i.e. by measuring the amount of gene product in its nucleotide form, any technology commonly used by the skilled person can be implemented. Methods for analyzing the level of expression of genes at the nucleotide level, such as, for example, transcripton analysis, include well-known methods such as RT-PCR or quantitative RT-PCR or nucleic acid chips. . "Nucleic acid chips" here means several different nucleic acid probes that are attached to a substrate, which may be a microchip, a glass slide, or a microsphere - sized bead. The microchip may be polymers, plastics, resins, polysaccharides, silica or a material based on silica, carbon, metals, inorganic glass, or nitrocellulose. The probes can be nucleic acids such as cDNAs ("cDNA chip"), RNAnn ("RNAnn" chips) or oligonucleotides ("oligonucleotide chips"), said oligonucleotides typically being able to have a length of between about 25 and 60 nucleotides. To determine the expression profile of a particular gene, a nucleic acid corresponding to all or part of said gene is labeled and then brought into contact with the chip under hybridization conditions, leading to the formation of complexes between said nucleic acid. labeled target and probes attached to the surface of the chip that are complementary to this nucleic acid. The presence of labeled hybridized complexes is then detected.

Preferably, the invention is carried out using any current or future method for determining gene expression based on the amount of RNAnn in the sample. For example, one skilled in the art can measure the expression of a gene by hybridization with a labeled nucleic acid probe, for example by Northern blot (for RNAn) or by Southern blot (for cDNA) but also by techniques such as the Serial Analysis Method of Gene Expression (SAGE) and its derivatives, such as LongSAGE, SuperSAGE, DeepSAGE, etc. It is also possible to use fabric chips (also known as TMAs: - tissue nnicroarrays "). The tests usually employed with tissue chips include immunohistochina and fluorescent in situ hybridization. For nRNA analysis, the tissue chips can be coupled with fluorescent in situ hybridization. Finally, it is possible to use bulk sequencing in parallel to determine the amount of RNAnn in the sample (RNA-Seq or Whole Transcripton Shotgun Sequencing). For this purpose, several methods of mass sequencing in parallel are available. Such methods are described in, for example, US 4,882,127, U.S. 4,849,077; U.S. 7,556,922; U.S. 6,723,513; WO 03/066896; WO 2007/111924; US 2008/0020392; WO 2006/084132; US 2009/0186349; US 2009/0181860; US 2009/0181385; US 2006/0275782; EP-B1-1141399; Shendure And Ji, Nat Biotechnol., 26 (10): 1135-45. 2008; Pihlak et al., Nat Biotechnol., 26 (6): 676-684, 2008; Fuller et al., Nature Biotechnol., 27 (11): 1013-1023, 2009; Tuesdays, Genonne Med., 1 (4): 40, 2009; Metzker, Nature Rev. Genet., 11 (1): 31-46, 2010. When the biomarker is a gene, and the expression level of the marker is measured at the peptide level, i.e. by measuring the amount of product of the gene in its peptide form, any method for determining the level of expression of a polypeptide known to those skilled in the art can be used. The methods for determining the level of expression of a polypeptide include, for example, mass spectrometry, biochemical tests, including immunological tests such as, for example, standard detection immunoassays (ELISA and ELISPOTS tests), or alternatively, for example, immunological tests using supported protein transfer techniques, such as the slot blot (also called dot blot) or the western blot. For example, it is possible to use protein chips, antibody chips, or tissue chips coupled to immunohistochina. Other techniques that can be used include FRET or BRET techniques, microscopy or histochemistry methods, including confocal microscopy and electron microscopy, methods based on the use of one or more several wavelengths of excitation and a suitable optical method, such as an electrochemical method (voltannnnetry and annperation techniques), the atomic force microscope, and radiofrequency methods, such as multipole resonance spectroscopy, confocal and non-confocal, fluorescence detection, luminescence, chemiluminescence, absorbance, reflectance, transnittance, and birefringence or refractive index (for example, by surface plasmon resonance, or - plasmon resonance surface in English, by ellipsonnetry, by method of resonant mirror, tec.), flow cytonetry, radioisotopic or magnetic resonance imaging, analysis by polyacrylannide gel electrophoresis (SDS-PAGE); by HPLC-Mass spectrophotometry, by liquid chromatography / mass spectrophotometry / mass spectrometry (LC-MS / MS). All these techniques are well known to those skilled in the art and it is not necessary to detail them here. According to a preferred embodiment, the level of expression of the marker chosen from desnnylin 1, involucrin, or filaggrin in its peptide form is measured by ELISA. An example of an antibody that can be used for an ELISA test that is suitable for measuring the level of expression of desnoglein 1 at the peptide level is the polyclonal rabbit antibody against the desnnogenin marketed by Abcann (ref. Ab14417). An example of an antibody usable for an ELISA test suitable for measuring the level of expression of involucrine at the peptide level is the mouse monoclonal antibody against the involucrin marketed by Sigma (ref 19018). An example of an antibody that can be used for an ELISA test suitable for measuring the level of expression of filaggrin at the peptide level is the rabbit polyclonal antibody against filaggrin marketed by Abcann (ref. Ab81468).

When the biological marker is a lipid, a sugar or a metabolite, a person skilled in the art may use any measurement method allowing said marker to be assayed in the sample of the subject. These methods include, for example, mass spectrometry and biochemical tests. According to a preferred embodiment, the level of expression of hyaluronic acid is measured by competitive ELISA. The competitive ELISA test of hyaluronic acid is a competitive assay that uses a protein that binds specifically to labeled (eg, biotinylated) hyaluronic acid, i.e., capable of binding to hyaluronic acid with high affinity, and microplates coated with hyaluronic acid. The labeled binding protein binds to the immobilized hyaluronic acid if it is not saturated with the free hyaluronic acid contained in the sample. During the next incubation, a conjugate capable of binding to the marker of the labeled protein (e.g. streptavidin peroxidase-conjugated conjugate) binds to the labeled binding protein. The reaction of the substrate is measured by the intensity of the color, which is inversely proportional to the level of hyaluronic acid contained in the samples. An example of a protein that binds specifically to hyaluronic acid (HA detector) and antibodies that can be used for a competition ELISA test that is suitable for measuring the level of expression of hyaluronic acid are those of the Tebu kit (ref. K-1200-0001). Lipids, in particular ceranides, may for example be assayed by conventional methods for measuring the incorporation of radionucleated acetate into reconstructed keratinocytes, or epidermis, or skin explants.

For the purposes of this application, the term "reference skin cell sample" means any sample containing human skin cells used as a reference sample. The person skilled in the art will be able to choose the reference skin sample according to, for example, the age, sex and skin color of the test subject.

Preferably, according to the invention, the sample of cutaneous reference cells is a sample of cutaneous cells of a child. In a particularly preferred manner, the reference skin sample is a skin sample of a child 2 years of age or older, an infant or a newborn.

The person skilled in the art may furthermore choose, as sample of reference skin cells, a sample of cells which have not been treated, or which have been irradiated, for example by UV. Preferably, when the biological marker according to the invention is claudin 1, or when the combination of biological markers of the invention comprises claudin 1, the sample of cutaneous reference cells is a sample of cells which have been irradiated with UV light. . According to a particular embodiment, the sample of cutaneous cells of reference according to the invention is a sample of cutaneous cells of said subject not having been treated by said active agent. Alternatively, according to another particular embodiment, the sample of cutaneous reference cells according to the invention is a sample of cutaneous cells of a subject having said deficiency of the cutaneous barrier, said subject having not been treated for said deficiency. According to a particular embodiment, the sample of cutaneous reference cells according to the invention is a sample of cutaneous cells of a healthy subject. Those skilled in the art will readily understand that the comparison of step c) is preferably performed between expression level measurements obtained for cutaneous cell samples that are of similar size, volume, or weight. Thus, it is preferable that the size, and / or the volume and / or the weight of the sample of step a) does not differ by more than 5% of the size, and / or the volume and / or the sample weight of step b). Even more preferably, the size, and the volume and the weight of the sample of step a) does not differ by more than 5% from the size, the volume of the sample weight of step b). Alternatively, if the samples of steps a) and b) differ by more than 5% by size, and / or volume and / or weight, those skilled in the art can normalize the levels obtained in steps a) and b). ) using a normalization factor. This factor may for example be a directly accessible physical marker such as the mass of cells of the sample, or the mass of a cellular constituent, such as the mass of cellular DNA or the mass of cellular proteins. It may also be advantageous to use as a normalizing factor the level of expression of a gene that is expressed at the same level in all or substantially all cells of the body. In other words, according to a particular embodiment of the present invention, the level of expression of a household gene is used as normalization factor. According to another embodiment, the levels of steps a) and b) are normalized using the level of expression, not of the housekeeping genes, but of the proteins encoded by them. A household gene is a gene expressed in all cell types, which encodes a protein having a basic function necessary for the survival of all cell types. A list of human housekeeping genes can be found in Eisenberg et al. (Trends in Genetics 19: 362-365, 2003). The household genes according to the invention include, for example, the genes B2M, TFRC, YWHAZ, RPLO, 185, GUSB, UBC, TBP, GAPDH, PPIA, POLR2A, ACTB, PGK1, HPRT1, IP08 and HMBS. Those skilled in the art can thus easily evaluate the effectiveness of the asset of the invention as a function of the comparison of step c). For example, when the level of desnnylin 1, involucrin, filaggrin, claudin 1 and / or hyaluronic acid, in particular in their peptide form, measured in step a), is greater than or equal to the level of measured in step b) in which said reference skin cell sample is a sample of cutaneous cells of a healthy subject, preferably a sample of cutaneous cells of a child, a child of 2 years or more, of an infant or a newborn, then the active agent is effective in the prevention and / or treatment of at least one deficiency of the cutaneous barrier of a subject. Likewise, for example, when the level of desnnylin 1, involucrin, ceranide 1, filaggrin, PAD1, hyaluronic acid, transglutaminase 1 and / or claudin 1, measured in step a), is greater than the level of the same marker measured in step b) in which said reference skin cell sample is a sample of cutaneous cells of said subject not having been treated by said active agent, or of a subject exhibiting said deficiency of the skin barrier and having not been treated for said deficiency, then the active agent is effective in the prevention and / or treatment of at least one deficiency of the cutaneous barrier of a subject.

For example, when the level of desnnylin 1, involucrin, ceranide 1, filaggrin, PAD1, hyaluronic acid, transglutaminase 1 and / or claudin 1 measured in step a), is less than or equal to at the level of the same marker measured in step b) in which said reference skin cell sample is a sample of cutaneous cells of said subject not having been treated by said active agent, or of a subject presenting said barrier deficiency cutaneous and not treated for said deficiency, then the asset is not effective in the prevention and / or treatment of at least one deficiency of the cutaneous barrier of a subject. By "deficiency of the cutaneous barrier" is meant within the meaning of the present application one or physiological or pathological modifications of the cutaneous barrier, more particularly one or physiological or pathological changes in the structure and / or function of the stratum corneum. and / or stratum granulosum, in comparison with normal adult skin. For the purposes of the present application, the deficiency of the cutaneous barrier may be due to a physiological modification of the cutaneous barrier. Children's skins, for example, even when healthy, may present a barrier that is not fully functional compared to adult skin. Indeed, the maturation of the child's skin is accompanied by a development of the barrier function thereof. In the same way, the skins of the elderly also show changes in the cutaneous barrier compared to the normal skin of adults, because of their senescence. For the purposes of the present application, the skins of the children and the skins of the elderly subjects have at least one deficiency of the cutaneous barrier. The immaturity of the skin of the children, and the senescence of the skin of the elderly are considered as deficiencies of the cutaneous barrier within the meaning of the present application. In addition, the deficiency of the cutaneous barrier can be due to a pathological modification of the cutaneous barrier. For example, skin damaged by external factors such as climatic conditions, pollution, injuries (eg cuts or burns), chemical irritations have decreases in the skin barrier.

Moreover, certain dermatological pathologies are associated with weakening of the cutaneous barrier, such as, for example, infant acne, adolescent acne, rosacea or erythrocouperosis, psoriasis, dermatitis of the seat, atopic dermatitis. , eczema, contact dermatitis, irritative dermatitis and in particular irritative dermatitis of the seat or diaper rash, allergic dermatitis, seborrheic dermatitis, dry skin, hyperreactivity of the skin. Preferentially, the deficiency of the cutaneous barrier is chosen from infant acne, adolescent acne, rosacea or erythrocouperosis, psoriasis, seat dermatitis, atopic dermatitis, eczema, contact dermatitis. irritative dermatitis and in particular irritative dermatitis of the seat or diaper rash, allergic dermatitis, seborrheic dermatitis, sensitive skin, reactive skin, xerosis, dehydrated skin, sun-damaged skin, radiation, by the wind, by the cold, by the heat, by the stress, by the pollution, the erythema cutaneous, the aged skin or photo aged, the skin photosensibilité, the scars, the ichthyoses, the cracks, the burns, the blows sun, inflammations due to rays of all kinds, irritation by chemical agents, physical, bacteriological, fungal or viral, parasitic. The method of the invention makes it possible to evaluate the effectiveness of an active agent chosen from C7 sugars and derivatives of formula (I), CH2ORi Ra-C-Rb R30 H R40 HH OR6 H OR6 CH2OR7 (I) where Ra represents a hydrogen atom and Rb represents a -OR2 or CRaRb represents the radical CO; R 1, R 2, R 3, R 4, R 5, R 6 and R 7 represent, independently of one another - a hydrogen atom or a - (CO) -R radical in which R represents a saturated or unsaturated hydrocarbon chain containing 11 to 24 carbon atoms, optionally substituted with one or more substituents selected from the group consisting of hydroxy (-OH) radicals, ethoxy radicals (-OC2H5) and -503M group with M representing a hydrogen atom, an ion ammonium NH4 + or a metal ion; or a - (CO) -R 'radical in which R' represents a saturated or unsaturated hydrocarbon chain containing from 2 to 10 carbon atoms, optionally substituted with one or more substituents chosen from the group consisting of hydroxyl (-OH) radicals; , ethoxy radicals (-OC2H5) and -503M group with M representing a hydrogen atom, an NH4 + ammonium ion or a metal ion. According to one particular embodiment of the invention, the active ingredient is phenolheptulose, of general formula (II) ## STR1 ## According to another particular embodiment of the invention, the active ingredient is is perseitol, of general formula (III) The source of C7 sugars, in particular of D-aminotheptulose and / or perseitol, may be a water-soluble extract of sugars avocado or other plant. Otherwise, D-Mannoheptulose and Perseitol are commercially available (synthetic origin). According to an advantageous variant of the invention, the source of D-rnannoheptulose and / or perseitol is a water-soluble extract of avocado sugars. The water soluble extract of avocado sugars can be directly obtained from any part of the avocado or avocado, such as the fruit, skin, kernel, leaf or roots of the avocado. avocado. It is also possible to obtain a water-soluble extract of sugars from co-products of the avocado processing industry, among which may be mentioned in a non-exhaustive way: the fresh avocado pulp, the frozen pulp, dehydrated, avocado cakes from oil extraction processes (mechanical extraction and / or solvent of the previously dehydrated fruit), deoiled solids from wet oil extraction processes (process of centrifugation), the deoiled solids from enzymatic avocado oil extraction processes, raw avocado purees (guacarnole), solid waste from the production units of these purees.

The extract is advantageously obtained from the fresh fruit of the avocado tree. The fruits can be selected from the varieties Hass, Fuerte, Ettinger, Bacon, Nabal, Anaheim, Lula, Reed, Zutano, Queen, Criola Selva, Mexicana Canta, Dschang Region, Hall, Booth, Peterson, Collinson Redn, more advantageously among the Hass, Fuerte and Reed varieties. Preferably, the Hass, Fuerte, Ettinger and Bacon varieties, and more preferably the Hass and Fuerte varieties, are preferred. In order to obtain C7 sugars, in particular D-nannoheptulose and / or perseitol, any method making it possible to obtain plant osidic extracts known to those skilled in the art may be used. Those skilled in the art may in particular refer to the process described in application FR 2 843 027. According to one embodiment of the invention, the C7 sugars, in particular Dnnannoheptulose and perseitol, are at least partially esterified with a radical - (CO) -R wherein R represents a saturated or unsaturated hydrocarbon chain containing from 11 to 24 carbon atoms, optionally substituted with one or more substituents selected from the group consisting of hydroxy (-OH) radicals, ethoxy radicals (-OC2H5) and -503M group with M representing a hydrogen atom, an NH4 + ammonium ion or a metal ion. In particular, the C7 sugars are at least partially esterified with a fatty acid residue. The hydrocarbon chain may be linear or branched, it is advantageously linear. It will be obvious to those skilled in the art that the assets can be formulated to facilitate administration, for example. According to one embodiment of the invention, the active ingredient is formulated in the form of a composition, preferably adapted for topical administration. The composition may further comprise at least one other active compound in addition to C7 sugars and derivatives of formula (I). This other compound may be chosen from all the compounds and their functional equivalents, stated below: This other compound may in particular be chosen from active agents conventionally used in dermatology or cosmetics such as emollients, moisturizing active agents, keratin synthesis, kératorégulateurs, keratolytiques, skin barrier restructuring agents, PPAR agonists (or Peroxysonne Proliferator Activated Receptor), RXR or LXR agonists, cicatrizing agents, sebo-regulating agents, anti-aging agents, irritating agents, soothing agents, anti-inflammatory agents, anti-oxidants and anti-aging agents, depicting or hypodecreating agents, pigmenting agents, lipolytic agents or lipogenesis inhibitors or anti-cellulite agents or slimming, mineral or organic filters and sunscreens, antifungal compounds, preservatives, antibacterial agents, pre and probiotics, antibiotics, immunomodulators. The invention is particularly useful for monitoring the progress of subjects, depending on the treatment administered and can therefore be used by the practitioner to choose the best treatment to administer to the subject. The subject of the invention is a method for adapting the treatment of a subject, said treatment comprising an active agent chosen from C7 sugars and derivatives of formula (I), ## STR2 ## Wherein Ra represents a hydrogen atom and Rb represents a -OR2 or CRaRb represents the CO radical; R1, R2, R3, R4, R5, R6 and R7 represent, independently of one another, a hydrogen atom or a - (CO) -R radical in which R represents a saturated or unsaturated hydrocarbon chain containing 11 to 24 carbon atoms, optionally substituted with one or more substituents selected from the group consisting of hydroxy (-OH), ethoxy (-OC2H5) and -503M with M representing hydrogen, ammonium ion NH4 + or a metal ion; or a - (CO) -R 'radical in which R' represents a saturated or unsaturated hydrocarbon chain containing from 2 to 10 carbon atoms, optionally substituted with one or more substituents chosen from the group consisting of hydroxyl (-OH) radicals; ethoxy radicals (-OC2H5) and -503M group with M representing a hydrogen atom, an NH4 + ammonium ion or a metal ion; said method comprising the steps of a) evaluating the effectiveness of said asset by a method for evaluating the effectiveness of an asset according to the invention; b) adapting said treatment according to the results obtained in step a).

Those skilled in the art will readily understand that the method of the invention for adapting the treatment of a subject can be implemented for the various assets mentioned above, as well as for the compositions comprising them. According to one particular embodiment of the invention, the active ingredient is phenolheptulose, of general formula (II) ## STR1 ## According to another particular embodiment of the invention, the active ingredient is is perseitol, of general formula (III) ## STR1 ## Those skilled in the art can easily adapt said treatment according to the results obtained in step a). For example, if the evaluation of step a) shows that the asset is not effective in the treatment and / or prevention of at least one deficiency of the barrier, the skilled person can consequently increase the quantities of assets administered, the concentration in assets or the frequency of administrations. Alternatively, the skilled person may for example adapt the treatment by adding other complementary agents of perseose avocado.

If, on the other hand, the active agent is effective, as determined in step a, in the treatment and / or prevention of at least one deficiency of the barrier, the person skilled in the art may, for example, continue the treatment without modify the quantities of assets administered, the concentration in assets or the frequency of administrations, reduce the quantities of assets administered, the concentration in assets or the frequency of administrations, or discontinue treatment. The method of the invention is therefore a practical tool and a decision aid for the skilled person. The third object of the invention is a kit for carrying out a method according to one of claims 1 to 11, comprising the means necessary for measuring the level of expression and / or activation of at least a biological marker chosen from desnnylin 1, involucrin, ceranides, in particular from ceranides 1 to 9, even more particularly ceranide 1, filaggrin, PAD1, hyaluronic acid, transglutaminase 1, and claudines, in particular claudin 1. According to a particular embodiment, the kit according to the invention comprises the means necessary for measuring the level of expression and / or activation of each of the biological markers of the combination consisting of desnnylin 1 , involucrin, ceranide 1, filaggrin, PAD1, hyaluronic acid, transglutaminase 1 and claudin 1. Preferably, the means necessary for measuring the level of expression of desnnylin 1, involucrin , filaggrin, claudin 1 or hyaluronic acid are antibodies specific for said markers. Preferably, the means necessary for measuring the expression level of PAD1 comprise nucleic probes and / or amplification primers capable of binding to PAD1 in its nucleotide form.

Legends of Figures Figure 1: Levels of expression of the involucrine marker in normal human keratinocytes, with or without treatment with perseose avocado (perseose). Figure 2: Levels of expression of the n-genin marker in normal human keratinocytes, with or without treatment with avocado perseose (perseose). Figure 3: Levels of transglutanninase marker activity in normal human keratinocytes, with or without treatment with avocado perseose. Figure 4: Expression levels of the filaggrin marker in normal human keratinocytes, with or without treatment with perseose avocado.

Figure 5: Levels of expression of the PAD1 marker in normal human keratinocytes, with or without treatment with perseose avocado. Figure 6: Levels of expression of the hyaluronic acid marker in normal human keratinocytes, with or without treatment with perseose avocado.

The examples which follow illustrate the invention but are not limiting. EXAMPLES 1. EXAMPLE 1 Preparation of a Water-Soluble Extract of Avocado Sugars The fresh avocados, of the Hass variety, are cut into thin strips of 2 to 5 mm thick, including the core, using a disc slicer. The drying tool is a thermoregulated oven with hot air flow. The sliced avocados are spread over a thickness of 4 to 5 cm on tiered shelves. The drying temperature is set at 80 ° C, the duration is 48 hours. Once dried, the fruits are under cold pressure. This operation is performed on a small laboratory Konnet press. Thus are obtained oil and a cake. The cake is then ground and extracted, in the presence of 70% ethanol or water. The liquid and solid parts are separated by centrifugation, for example. The soluble (liquid) fraction is taken up again to be purified and concentrated according to the following procedure: - Demineralization using ion exchange resins: demineralization of the heptuloses by passage on OH- resins, then on H + resin. Ultrafiltration over 10,000 Da: ultrafiltration is carried out with a system equipped with 4 membranes of 10 kDa cut-off threshold. - Concentration under vacuum: the concentration of the purified extract is carried out using a vacuum evaporator to obtain a dry matter close to 4%. Packaging: the concentration of the extract is adjusted to 5% of dry matter and preservative is added, followed by sterile filtration with a membrane of 0.2% cut-off point and conditioned. Table 1 gives the composition of the C7 avocado sugar extract, at 5% dry matter, prepared according to the method described above: Appearance Pale yellow solution Analytical criteria Dry matter 5% pH (dilution 1 / 4) 7.0 Composition (% / dry matter) Sucrose 3.0 Glucose 7.5 D-Annoheptulose 40.0 Fructose 8.6 Perseitol 40.0 Table 1 Following the same procedure, two other extracts were prepared, of which the pH value, the absorbance and the C7 sugar content are given in Table 2. The C7 sugar content corresponds to the sum of perseitol and D-methyloheptulose analyzed by HPLC. Lot 1 2 Dry matter 5% 5% pH (1/4 dilution) 5.9 5.4 Sugars in C7 / dry matter 80.5 83.4 Table 2 Avocado sugars prepared by the process of Example 1 and called - avocado perseoses "below can be used in different formulations.

Examples of formulations Moisturizing cream Raw material / Name 1 to 15% commercial CAPRYLO CAPRATE GLYC SUNFLOWER OIL 1 to 15% PURE CETYLIC ALCOHOL 1 to 5% GLYCERYL STEARATE CITRATE 1 to 10% 1 to 5%,, 1 BEE WAX, I - [-É-UMULGINSG I, 0 to 2%,, 1, 1 I ACETATE VITAMIN E, 0 to 1% 1 1 PURIFIED WATER QSP 100% I CARBOPOL ULTREZ20 0 to 1% GLYCEROL 1 to 10% XANTHANE GUM 0 to 1% LAUNDRY 0 to 1% PRESERVATIVE 0 to 2% PERSEOSE LAYER 0 to 1% Table 35 Restructuring milk Raw material / Name WATER PURIFIED QSP 100% SUNFLOWER OIL 1 to 10% OIL OF COPRAH HYDRO 1 to 10% OIL OF SOFT ALMOND 1MARKET OIL MONOSTEARATE GLYCEROL STEARIC ACID CONSERVATIVE CETYLIC ALCOHOL C16-C18 ACETATE VITAMIN E LAUNDRY SOUDEXI PERSEOSE AVOCATE 10% to 10% to 10% to 10% 0 to 2% Table 4 Cleansing water Raw material / Name WATER PU RI Fl EE QSP 100% GLYCEROL 1 to 10% SODIUM COCOYL 1 to 10% CONSERVATIVE 0 to 2% RICIN HYDROBUTETH26 0 to 2% ALLANTOIN 0 to 2% ACID TARTRIQ EU 0 to 2% ALOE VERA POWDER 0 to 2% SAPONARY EXTRACT 0 to 2% LAUNDRY 0 to 2% LESS CONTROL 0 to 1% Table 5 Shampoo Raw Material / Name PURIFIED WATER of QSP 100% COCAMIDO PROPYL BETAINE 1 to 10% GLYCEROL 1 to 10% COCOGLUCOSIDEXI 1 to 10% SODIUM MYRETH SULFATE 1 to 10% PRESERVATIVE 0 to 2% DISTEARATEPEG6000 0 to 2% POLYQUARTE RN IUMJR400X1 0 to 2% PANTHENOL DEXTROGYRE 0 to 2% CITRIC ACID HYDXI 0 to 2% PERSEOSE AVOCADO 0 at 1% Table 6 Sun cream SPF 50+ Raw material / Trade name CAPRYLATE / CAPRATEDECOPRAH 5 to 20% I DICAPRYLYLCARBONATE 5 to 20% CAPRYLOCAPRATEGLYC 5 to 20% MIGLYOLGELB 5 to 20% TITANIDIOXIDJOJOBAESTERS 1 to 15% TINOSORBS 1 to 5% DIETAMIN HYDBENZO YHEXBENZ 1 to 10% ETHYL HEXYL TRIAZONE 1 to 10% HAREF AVIATION OIL 1 to 5% ALPHA TOCOPHEROL 0.05 to 1% LAURYL GLUCOSE GLYSTEARATXI 2 to 12% PURIFIED WATER QSP 100% 1GLYCEROL 1 to 10% XANTHAN GUM 0 at 1% CONSERVATIVE 0 to 2% POTASSIUM CETYL PHOSPHATE 0 to 2% LAUNDRY SOUDEXI 0 to 2% P HENYL BENZIMIDAZO SULFONAC 1 to 5% PERSEOSE LAWYER 0 to 1% Table 7 The avocado sugars prepared by the method of Example 1 and called "avocado perseose" below were used to study the expression of the markers of the barrier function under different conditions. 2. Epidermal maturation a. Expression of Involucin Material and Methods: Normal human epidermal keratinocytes were incubated in the presence of 0.005% and 0.05% (w / v) avocado perseose for 48 hours.

At the end of the treatment, the involucrine produced was assayed on the surface of the keratinocytes by a cell ELISA technique. The amount of involucrine was referred to the number of live cells estimated by a MU assay. Results: The results are shown in Table 8 below, and in Figure 1.

Persarose avocado at 0.005% and 0.05% significantly stimulated the expression of involucrine in normal human keratinocytes: + 17%, p <0.01 and + 56%, respectively, p <0.001. Involucrine (DOELISAIDOMTT) Control cells 2,245 ± 0,068 Perseose of avocado 0,005% 2,633 ± 0,051 + 17% ** Perseose of avocado 0,05% 3,509 ± 0,178 + 56% *** Table 8 ** p <0,01; *** p <0.001 vs control cells, one-way analysis of variance followed by a Dunnett test (GraphPad Prisnn software) b. Desnnyline Expression Material and Methods: Normal human epidermal keratinocytes were cultured in a differentiating inducing medium (supplemented with Ca ++) and incubated in the presence of 0.005% and 0.05% avocado perseose for 48 hours. At the end of the treatment, the desnnyline-1 produced was measured on the surface of the keratinocytes by an ELISA technique on cells. The amount of desnnolinin was related to the number of living cells estimated by a MU assay. Results: The results are shown in Table 9 below, and in Figure 2. 0.005% and 0.05% avocado perseose significantly stimulated the expression of desnnylin 1 in normalized human keratinocytes differentiated: Respectively + 41%, p <0.001 and + 40%, p <0.001. Desmoglein (DOELISAIDOMTT) Control cells 5,791 ± 0,641 Avocado perseose 0,005% 8,182 ± 0,430 + 41% *** Perseose of avocado 0,05% 8,094 ± 0,395 + 40% *** Table 9 *** p <0,001 vs control cells, one-way analysis of variance followed by a Dunnett test (GraphPad Prisnn software) 3. Reinforcement of the lipid barrier: neosynthesis of ceranides Material and methods: Epidermis reconstructed on day 5 were cultured under the following conditions: Epidermal Controls: Depleted Medium, Differentiated Epidermis: Complete Differentiation Medium, Persarent Avocado at 0.005% and 0.05% (MS) in the Depleted Medium. After 24 hours of incubation, the culture medium was renewed and supplemented with radio-labeled acetate (14C-acetate). The epidermis was then incubated again for 48 hours.

At the end of the treatment, the neo-synthesized ceranides were analyzed by quantification of the radioactivity incorporated after thin layer chromatography. Results: The results are shown in Table 10 below. Perseose of avocado at 0.005% and 0.05% increased the synthesis of ceranides in reconstructed human epidermis: respectively + 32% and + 27%. Amount of Ceramide Relative Intensity% of control (average) Control Epiderms 14.2 100 Differentiated Epidermis 19.7 138 Avocado Perseosis 0.005% 18.0 132 Avocado Perseosis 0.05% 18.7 127 Table 10 4. Regulation hydric a. Enzymatic Activity of Transglutaminase Material and Methods: Normal human epidermal keratinocytes were incubated in the presence of 0.005% and 0.05% avocado perseose in KGM Gold medium (Lonza); a control of cells grown in a pro-differentiating medium (KGM Gold supplemented with Ca ++) was carried out in parallel. After 72 hours of incubation, the cells were lysed and the intracellular transglutanninase activity assayed using the Transglutanninase Colorinnetric Microassay Kit (Covalab). The results were normalized to the amount of total protein determined using the BC Assay kit (Interchinn). Results: The results are shown in Table 11 below, and in Figure 3.

0.005% avocado perseose significantly stimulated the enzymatic activity of transglutaminase in normal human epidermal keratinocytes: + 175%, p <0.05. Transglutaminase activity (OD / total protein) Control cells 0.000012 ± 0.0000018 Reference (Ca) 0.00003 ± 0.0000054 + 146% * Perseosis of avocado 0.005% 0.000033 ± 0.0000075 + 175% * Perseosis of avocado 0.05% 0.000025 ± 0.0000086 + 110% ns Table 11 * p <0.05 vs control cells, one-way analysis of variance followed by a Dunnett test (GraphPad Prisnn software) b. Expression of Filaggrin Material and Methods: Normal human epidermal keratinocytes were incubated in the presence of 0.005% and 0.05% avocado perseose for 72 hours.

At the end of the treatment, the filaggrin produced was measured on the surface of the keratinocytes by a cell ELISA technique. The amount of filaggrin was related to the number of living cells estimated by a Neutral Red assay. Results: The results are shown in Table 12 below, and in Figure 4. Persian avocado at 0.05% significantly stimulated the production of filaggrin by normal human epidermal keratinocytes: + 195%, p < 0.01.

Filaggrin (DOELISA 1 DORN) Control cells 0.683 ± 0.089 Avocado perseose 0.005% 1.102 ± 0.681 + 61% ns Perseosis of avocado 0.05% 2014 ± 0.546 + 195% ** Table 12 ** p <0.01 vs control cells, one-way analysis of variance followed by a Dunnett test (GraphPad Prisnn software). vs. Gene Expression of PAD1 Enzyme (Peptidyl Arginin Deinninase) Materials and Methods: Normal human epidermal keratinocytes cultured in a differentiation-inducing medium (supplemented with Ca ++) were incubated for 48 hours in the presence of 0.005 Persian avocado. % and 0.05% or vitamin D3 at 10-7M (1a, 25-Dihydroxyvitannin D3; Sigma), positive reference.

At the end of the treatment, the RNAs were extracted and the gene expression of PAD1 was evaluated by RT-PCR in real time, the GAPDH reference gene was used to normalize the results. Results: The results are shown in Table 13 below, and in Figure 5.

Persarose avocado 0.05% significantly stimulated gene expression of PAD1: + 118%, p <0.05. PAD1 (Relative Quantity standardized by GAPDH) Control cells 1.00 Vitamin D3 10-7M 2.66 + 166% ** Perseose of avocado 0.005% 1.27 + 27% ns Perseosis of avocado 0.05% 2.18 + 118% * Table 13 * p <0.05; ** p <0.01 vs control cells, one-way analysis of variance followed by a Dunnett test (GraphPad Prisnn software) d. Production of Hyaluronic Acid Material and Methods: Normal human epidermal keratinocytes were incubated in the presence of 0.005% and 0.05% avocado persecrosis or 10-7M retinoic acid (all-transRetinoic acid; Sigma), positive reference. After 24 hours of incubation, the hyaluronic acid produced by the keratinocytes was assayed in the culture supernatants by an ELISA (Tebu) method. The amount of hyaluronic acid measured was related to the number of living cells estimated by a Neutral Red test. Results: The results are shown in Table 14 below, and in Figure 6. Perseose avocado significantly increased the amount of hyaluronic acid produced by normal human keratinocytes: + 47%, p <0.05 at 0.005% and + 43%, p <0.05 to 0.05%. Hyaluronic acid (ng / ml / Living cells) Control cells 248,445 ± 21,248 Ac. retinoic 10-7M 544.024 ± 34.026 + 119% *** Perseose of avocado 365.737 ± 61.68 + 47% * 0.005% Perseose of avocado 356.179 ± 37.353 + 43% * 0.05% Table 14 * p <0, 05; *** p <0.001 vs control cells, one-way analysis of variance followed by a Dunnett test (GraphPad Prisnn software). 5. UV aggression model Certain environmental aggressions are harmful for the maturation of the epidermis and especially for the barrier function (Yamamoto T, Kurasawa M, Hattori T, Maeda T, Nakano H, Sasaki H. Relationship between expression of tight junction In this study, the results of this study are summarized in the following: Arch Dernnatol Res 2008; 300: 61-8; Valacchi et al., Cutaneous responses to environmental stressors. Ann NY Acad Sci 2012; 1271: 7581, 2012). MATERIALS AND METHODS: Normal human epidermal keratinocytes cultured in a differentiation-inducing medium (supplemented with Ca ++) were preincubated in the presence of 0.001% avocado perseose for 48 hours or at 0.005% and 0.05% during 24 hours. The keratinocytes were then irradiated in PBS with UVA + B at a dose of 36 kJ / nn2. The cells were then incubated again for 6 hours in the presence of avocado perseose. At the end of the treatment, the RNAs were extracted and the gene expressions of claudin 1, filaggrin and involucrin were evaluated by RT-PCR in real time. The results were normalized by amplification of a reference gene: 62-nnicroglobulin for 0.001% persreose avocado or HPRT assays for 0.005% and 0.05% avocado perseose trials. Results: The results are shown in Table 15 below. UV irradiation induced a significant decrease in the gene expression of the three barrier markers studied. Avocado perseose counterbalanced the effect of UV by significantly stimulating gene expression of claudin 1 (+ 131%, p <0.001 to 0.001% and 32%, p <0.01 to 0.05%). ), filaggrin (+ 22%, p <0.05 to 0.005% and + 27%, p <0.01 to 0.05%) as well as involucrin (+ 191%, p <0.01) to 0.001%) under UV.

Claudine 1 Filaggrine Involucrine Control cells 1.00 1.00 1.00 1.00 Irradiated control 0.39 -61% $$ 5 0.50 -50% $$ 5 0.56 -44% $$ 5 0.29 -71% $$ 5 UVA + B 36kJ / m2 Perseosis 0'90 + 131% *** / / +191% ** 0.85 avocado 0.001% Perseose / 0.60 + 20% ns 0.69 + 22% * / avocado 0.005% Perseose / 0.66 + 32% ** 0.71 + 27% ** / avocado 0.05% Table 15 $$ 5 p <0.001 vs. control cells * p <0.05; ** p <0.01; *** p <0.001 vs irradiated control One-way analysis of variance followed by a Tukey test (GraphPad Prisnn software).

Claims (15)

REVENDICATIONS1. A method for evaluating the efficacy of an active agent selected from C7 sugars and derivatives of formula (I), CH2ORi Ra-C-Rb R30 H R40 HH OR5 H OR6 CH2OR7 (I) wherein Ra represents a hydrogen atom and Rb is -OR2 or CRaRb is CO; R1, R2, R3, R4, R5, R6 and R7 represent, independently of one another, a hydrogen atom or a - (CO) -R radical in which R represents a saturated or unsaturated hydrocarbon chain containing 11 to 24 carbon atoms, optionally substituted with one or more substituents selected from the group consisting of hydroxy (-OH), ethoxy (-OC2H5) and -503M with M representing hydrogen, ammonium ion NH4 + or a metal ion; Or a - (CO) -R 'radical in which R' represents a saturated or unsaturated hydrocarbon chain containing from 2 to 10 carbon atoms, optionally substituted by one or more substituents selected from the group consisting of hydroxy (-OH) radicals; ethoxy radicals (-OC2H5) and -503M group with M representing a hydrogen atom, an NH4 + ammonium ion or a metal ion; in the prevention and / or treatment of at least one skin barrier deficiency of a subject, said method comprising the steps of: a. measuring the level of expression and / or activation of at least one biological marker in a sample of cutaneous cells of the subject, characterized in that said biological marker is chosen from: the markers of epidermal maturation, said marker epidermal maturation being preferably selected from desmoglein 1 and involucrin, or; The markers of the lipid barrier, said marker of the lipid barrier being preferably chosen from ceramides, in particular from ceramides 1 to 9, even more particularly ceramide 1, or the markers of the water regulation, said marker of the wherein the water repetition is preferably selected from filaggrin, PAD4, hyatonic acid and transglutaminase 1, or; the markers of the regulation of the stratum granulosum, said regulation marker of the stratum granutosum preferably being chosen from claudines, in particular claudin 1; b. measuring the level of expression and / or activation of said biological marker in a sample of cutaneous reference cells; vs. comparing the expression and / or activation levels obtained in step a) with cl: expression and / or activation levels obtained in step b); d. evaluating the effectiveness of said asset based on the comparison of step b). 2. Method according to claim 1, characterized in that step a) comprises measuring the level of expression and / or activation of a combination of biological markers from a sample of cutaneous cells of the subject, characterized in that said column of markers comprises: at least one marker of epidermal maturation, said marker of epidermal maturation being preferably selected from desmoglein 1 and teotucrine; at least one marker of the lipid barrier, said marker of the Ledic barrier being preferably chosen from ceramides, in particular from cereals T 9, even more particularly ceramide; and at least one water-regulating marker, said water-regulating marker being preferably selected from filaggrin, PAD1, hyaluronic acid and transglutaminase 1, at least one regulatory marker of the stratum granulosum, said marker of the regulating the stratum granulosum being preferably chosen from claudines, in particular claudin 1 3. Method according to claim 2, characterized in that the combination of biological markers consists of desmoglein 1, involucrine, ceramide 1, filaggrin, PAD1, hyaluronic acid, transglutaminase 1 and claudin 1. 4. Method according to one of claims 1 to 3, characterized in that said sample of cutaneous cells of reference is a sample of cutaneous cells of said subject that has not been treated with said active ingredient. 5. Method according to one of claims 1 to 3, characterized in that said reference skin sample is a sample of cutaneous cells of a subject having said deficiency of the cutaneous barrier, said subject having not been treated. for said disability. 6. Method according to one of claims 1 to 3, characterized in that said sample of cutaneous cells is a sample of cutaneous cells of a healthy subject. 7. Method according to one of claims 1 to 6, characterized in that the sample of cutaneous cells of reference is a sample of cutaneous cells of a child, preferably of a newborn, that is to say of a child whose age is between 0 and 1 months, an infant, that is to say a child whose age is between 1 month and 2 years, or a child child 2 years old or over, that is to say, a child whose age is between 2 years and 16 years. 8. Method according to one of claims 1 to 7, characterized in that said deficiencedde the cutaneous barrier is selected from infant acne, adolescent acne, rosacea or erythrocouperosis, psoriasis, dermatitis of the skin. atopic dermatitis, eczema, contact dermatitis, irritative dermatitis and in particular irritative dermatitis of the seat or diaper rash, allergic dermatitis, seborrheic dermatitis, sensitive skin, reactive skin, xerosis, dehydrated skin, skin damaged by the sun, radiation, wind, cold, heat, stress, pollution, skin rash, aged or photoaged skin, photosensitized skin, scars, ichthyosis, sunstrokes, burns, sunburns, inflammations due to rays of all kinds, irritations by chemical, physical, bacteriological, fungal or viral, parasitic agents. 9. Method according to one of claims 1 to 8, characterized in that the expressicri level of the marker chosen from desmoglétne 1, involucrine, or filaggrine, corresponds to the level of expression of said marker in its peptide form. the level of expression of PAN or eaudin 1 corresponds to the level of expression of said marker in its nucleotide form. 10. Method according to one of claims 1 to 9, characterized in that said active agent is D-mannoheptutose, of the general formula (II) CH2OH ## STR1 ## 11. Method according to one of claims 1 to 9, characterized in that said active ingredient is Rament, of general formula (1H) CH2OH - H OH H H OH CHOH (III) 12. Method according to one of claims 1 to 11, characterized in that said active agent is in the form of a composition, preferably adapted for topical administration. 13. The method of claim 12, characterized in that the composition may further comprise at least one other active compound in addition to C7 sugars and derivatives of formula (I). 14. Kit for carrying out a method according to one of claims 1 to 11, comprising the means necessary for measuring the level of expression and / or activation of at least one biological marker chosen from the desmoglein 1, involucrine, ceramides, in particular from ceramides 1 to 9, more particularly ceramide 1, filaggrin, PAD1, hyaluronic acid, transglutaminase 1, and claudines, in particular claudin 1. 15. Kit according to claim 10, characterized in that it comprises the means necessary for measuring the level of expression and / or activation of each of the biological markers of the combination of biological markers consisting of desnnylin 1, l involucrin, ceranide 1, filaggrin, PAD1, hyaluronic acid, transglutaminase 1 and claudin 1.